以蚕豆(Vicia faba)为材料, 利用激光共聚焦显微技术和分光光度技术, 结合药理学实验, 探讨硫化氢(hydrogen sulphide, H2S)和过氧化氢(hydrogen peroxide, H2O2)在茉莉酸(jasmonic acid, JA)调控气孔运动信号转导中的作用。结果表明, H2S合成抑制剂氨氧基乙酸(aminooxy acetic acid, AOA)、羟胺(hydroxylamine, NH2OH)、丙酮酸钾(potasium pyruvate, C3H3KO3) 和氨水(ammonia, NH3), H2O2 清除剂抗坏血酸(ascorbic acid, AsA), 合成抑制剂水杨羟肟酸 (salicylhydroxamic acid, SHAM)、二苯基碘(diphenylene iodonium, DPI)均可逆转JA诱导的气孔关闭效应。JA能够明显提高蚕豆叶片及保卫细胞中的H2O2水平、H2S含量和L-/D-半胱氨酸脱巯基酶活性; H2S合成抑制剂可抑制JA引起的叶片H2S含量的增加; 而H2O2清除剂则可减弱JA对H2S含量变化和L-/D-半胱氨酸脱巯基酶活性的诱导效应。以上结果表明H2S和H2O2均参与了JA诱导的蚕豆气孔关闭, 且H2S(主要由L-/D-半胱氨酸脱巯基酶合成)可能作为H2O2的下游组分参与调控这
一信号转导过程。
Pharmacological treatments combined with laser scanning confocal microscopy (LSCM) and spectrophotography were used to study the role of H2S and H2O2 in the signaling transduction during stomatal movement responding to jasmonic acid (JA) in Vicia faba. Inhibitors of H2S synthesis (aminooxy acetic acid, hydroxylamine, and potasium pyruvate + ammonia), the scavenger of H2O2 (ascorbic acid), and the inhibitors of H2O2 synthesis (salicylhydroxamic acid, diphenylene iodonium) all reduced JA-induced stomatal closure. Moreover, JA enhanced H2O2 and H2S levels and L-/D-cysteine desulfhydrase activity in leaves and guard cells. The inhibitors of L-/D-cysteine desulfhydrase diminished JA-induced H2S production in leaves. In addition, H2O2 scavenger decreased H2S level and L-/D-cysteine desulfhydrase activity induced by JA. Therefore, H2S and H2O2 are involved in the signal transduction pathway of JA-induced stomatal closure. L-/D-cysteine desulfhydrase-derived H2S may represent a novel downstream component of the H2O2 signaling cascade during JA-induced stomatal movement in V. faba.
安国勇, 宋纯鹏, 张骁, 荆艳彩, 阳冬梅, 黄美娟, 周培爱, 吴才宏 (2000). 过氧化氢对蚕豆气孔运动和质膜K+通道的影响. 植物生理学报 26, 458-464.
薄惠, 王棚涛, 董发才, 宋纯鹏 (2005). 拟南芥保卫细胞中茉莉酸甲酯诱导的H2O2 产生与MAPK 信号转导体系的可能关系. 植物生理学通讯 41, 439-443.
崔香环, 苗雨晨, 董发才, 宋纯鹏 (2001). 茉莉酸甲酯诱导气孔关闭的信号转导机制初探. 河南大学学报(自然科学版) 31, 75-78.
李东波, 肖朝霞, 刘灵霞, 王金成, 宋国力, 毕玉蓉(2010). 外源硫化氢对豌豆根尖及其边缘细胞的影响. 植物学报45, 354-362.
刘国华, 侯丽霞, 刘菁, 刘新, 王学臣 (2009). H2O2介导的NO合成参与乙烯诱导的拟南芥叶片气孔关闭. 自然科学进展 19, 841-851.
刘新, 李云, 孟繁霞, 张蜀秋 (2007). H+参与茉莉酸调控蚕豆气孔运动的信号转导. 植物生理学通讯 43, 245-249.
刘新, 石武良, 张蜀秋, 娄成后 (2005). 一氧化氮参与茉莉酸诱导蚕豆气孔关闭的信号转导. 科学通报 50, 453-458.
左照江, 张汝民, 高岩 (2009). 植物间挥发物信号的研究进展. 植物学报44, 245-252.
Angelini R, Tisi A, Rea G, Chen MM, Botta M, Federico R, Cona A (2008). Invovement of polyamine oxidase in wound healing. Plant Physiol 146, 162-177.
Biswa R A, Sarah M A (2008). Hormone interactions in stomatal function. Plant Mol Biol 69, 451-462.
Bloem E, Riemenschneider A, Volker J, Papenbrock J, Schmidt A, Salac I, Haneklaus S, Schnug E (2004). Sulphur supply and infection with Pyrenopeziza brassicae in?uence L-cysteine desulphydrase activity in Brassica napus L. J Exp Bot 55, 2305-2312.
Brennan TC (1977). Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol 59, 411-416.
Gadalla MM, Snyder SHJ (2010). Hydrogen sulfide as a gasotransmitter. Neurochem 113, 14-26.
Gehring CA, Irving HR, McConchie R, Parish RW (1997). Jasmonates induce intracellular alkalinizaton and closure of Paphiopedilum guard cells. Ann Bot 80, 485-489.
Gonugunta VK, Srivastava N, Raghavendra AS (2009). Cytosolic alkalinization is a common and early messenger preceding the production of ROS and NO during stomatal closure by variable signals, including abscisic acid, methyl jasmonate and chitosan. Plant Signal Behav 4, 561-564.
Huang DQ, Wu WR, Abrams SR, Cutler AJ (2008). The relationship of drought-related gene expression in Arabidopsis thaliana to hormonal and environmental factors. J Exp Bot 59, 2991-3007.
Kolla VA, Vavasseur A, Raghavendra AS (2007). Hydrogen peroxide production is an early event during bicarbonate induced stomatal closure in abaxial epidermis of Arabidopsis. Planta 225, 1421-1429.
Munemasa S, Oda K, Watanabe-Sugimoto M, NakamuraY, Shimoishi Y, Murata Y (2007). The coronatine-insensitive 1 mutation reveals the hormonal signaling interaction between abscisic acid and methyl jasmonate in Arabidopsis guard cells. Speci?c impairment of ion channel activation and second messenger production. Plant physiol 143, 1398-1407.
Papenbrock J, Riemenschneider A, Kamp A, Schulz-Vogt HN, Schmidt A (2007). Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants- from the field to the test tube and back. Plant Biol 9, 582-588.
Riemenschneider A, Nikiforova V, Hoefgen R, Luit J, Kok D, Papenbrock J (2005). Impact of elevated H2S on metabolite levels, activity of enzymes and expression of genes involved in cysteine metabolism. Plant Physiol Biochem 43, 473-483.
Sekiya J, Schmidt A, Wilson LG, Filner P (1982). Emission of hydrogen sulfide by leaf tissue in response to L-Cysteine. Plant Physiol 70, 430-436.
She XP, Song XG (2008). Carbon monoxide-induced stomatai closure involves generation of hydrogen peroxide in Vicia faba guard cell. J Integr Plant Biol 50, 1539-1548.
Suhita D, Raghavendra AS, Kwak JM, Vavasseur A (2004). Cytoplasmic alkalization precedes reactive oxygen species production during methyl jasmonate- and abscisic acid-induced stomatal closure. Plant Physiol 134, 1536-1545.
Underwood W, Melotto M, He SY (2007). Role of plant stomata in bacterial invasion. Cell Microbiol 9, 1621-1629.
Veal EA, Day AM, Morgan BA (2007). Hydrogen peroxide sensing and signaling. Mol Cell 26, 1-14.
Yong QC, Choo CH, Tan BH, Low CM, Bian JS (2010).Effect of hydrogen sulfide on intracellular calcium homeostasis in neuronal cells. Neurochem Int 56, 508-515.
Zhang H, Hu LY, Hu KD, He YD, Wang SH, Luo JP (2008). Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress. J Integr Plant Biol 50, 1518-1529.
Zhang H, Ye YK, Wang SH, Luo JP, Tang J, Ma DF (2009). Hydrogen sul?de counteracts chlorophyll loss in sweet potato seedling leaves and alleviates oxidative damage against osmotic stress. Plant Growth Regul 58, 243-250.
Zhang H, Jiao H, Jiang CX, Wang SH, Wei ZJ, Luo JP, Jones RL (2010a). Hydrogen sul?de protects soybean seedlings against drought-induced oxidative stress. Acta Physiol Plant. http://www.springerlink.com/content/m31j70735733x5u6/
Zhang H, Tan ZQ, Hu LY, Wang SH, Luo JP, Jones RL (2010b). Hydrogen sulfide alleviates aluminum toxicity in germinating wheat seedlings．J Integr Plant Biol 52, 556-567.
Zhang X, Dong FC, Gao JF, Song CP (2001). Hydrogen peroxide-induced changes in intracellular pH of guard cells precede stomatal closure. Cell Res 11, 37-43.